Workflow for PCR Amplification of Genomic DNA: dna extraction kit
1. Sample Collection
- Obtain tissue, blood, cultured cells, plant material, or other biological specimens.
- Ensure proper storage (e.g., fresh, frozen, or preserved) to prevent DNA degradation.
2. Genomic DNA Extraction
- Lyse cells/tissues (mechanical, enzymatic, or chemical lysis).
- Remove proteins and contaminants (e.g., with proteinase K, detergents, or magnetic beads).
- Purify genomic DNA using a kit, spin column, or phenol-chloroform extraction.
- Quantify and assess purity (spectrophotometry at 260/280 nm or fluorometric assays).
3. PCR Reaction Setup
-
Prepare PCR master mix containing:
- Template DNA (extracted genomic DNA).
- Primers (forward and reverse, sequence-specific).
- dNTPs (nucleotide triphosphates).
- Buffer & MgCl₂ (for optimal enzyme activity).
- Thermostable DNA polymerase (e.g., Taq, high-fidelity polymerases).
- Aliquot into PCR tubes or plates.
4. Thermal Cycling
-
Place in a thermocycler and run:
- Denaturation (95 °C): DNA strands separate.
- Annealing (50–65 °C): primers bind to target sequence.
- Extension (72 °C): DNA polymerase synthesizes new strands.
- Repeat 25–40 cycles for exponential amplification.
5. PCR Product Analysis
- Run PCR products on agarose gel electrophoresis.
- Stain with DNA dye (e.g., ethidium bromide or SYBR Safe).
- Visualize under UV or blue-light transillumination.
6. Downstream Applications
- Sequencing of amplified fragments.
- Cloning into vectors.
- Diagnostic assays (pathogen detection, genetic screening).
- Genotyping or mutation analysis.
DNA Extraction Solution is a fast and simple method for extracting PCR-ready genomic DNA from almost any sample for screening and genotyping applications
- Fast: 8 minute extraction protocol for most sample types
- Simple: No need of centrifugation steps or spin columns to help increase yields
- Automation-friendly: Procedure can be easily scaled to process hundreds of samples using automated workflows
- Safe: Uses only non-toxic reagents
- Many applications: suitable for genotyping, human identity testing, viral/microbial screening, and more.
- Available in:
- 5 mL (10 extractions)
- 50 mL (100 extractions)
- 1000 mL (2000 extractions)
Your Dynamic Snippet will be displayed here... This message is displayed because you did not provide both a filter and a template to use.